Spectral fingerprint diagnosis: Spatially independent analysis of biomarker patterns in homogeneous systems.

Summary

sf-PCR encodes three-dimensional fluorescence spectral fingerprints (peak position and intensity) to overcome spectral overlap in homogeneous multiplex PCR. A 10-plex model shows linear superimposability and decodability, with demonstrated potential in cancer and respiratory pathogen detection.

Key Findings

• Introduced spectral fingerprint PCR that uses 3D fluorescence fingerprints to resolve spectral overlap in homogeneous multiplex assays.
• Demonstrated linear superimposability and decodability in a 10-plex sf-PCR model.
• Showed clinical potential in cancer diagnostics and respiratory pathogen detection.

Clinical Implications

sf-PCR could streamline multiplex respiratory pathogen panels with fewer channels and simpler optics, supporting rapid syndromic testing in clinical and near-patient settings.

Limitations

• Clinical validation breadth and head-to-head comparisons with gold-standard multiplex assays are limited in this report.
• Performance under complex clinical matrices and high target burden needs further assessment.

Future Directions

Benchmark sf-PCR against established multiplex respiratory panels across diverse specimen types, and integrate with portable optics for point-of-care deployment.